Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes
This transformation procedure generates, with high efficiency (70–90%), hairy roots in cultivars, landraces and accessions of Phaseolus vulgaris (common bean) and other Phaseolus spp. Hairy roots rapidly develop after wounding young plantlets with Agrobacterium rhizogenes , at the cotyledon node, an...
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| Veröffentlicht in: | Nature protocols 2007-07, Vol.2 (7), p.1819-1824 |
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| creator | Estrada-Navarrete, Georgina Alvarado-Affantranger, Xochitl Olivares, Juan-Elías Guillén, Gabriel Díaz-Camino, Claudia Campos, Francisco Quinto, Carmen Gresshoff, Peter M Sanchez, Federico |
| description | This transformation procedure generates, with high efficiency (70–90%), hairy roots in cultivars, landraces and accessions of
Phaseolus vulgaris
(common bean) and other
Phaseolus
spp. Hairy roots rapidly develop after wounding young plantlets with
Agrobacterium rhizogenes
, at the cotyledon node, and keeping the plants in high-humidity conditions. Callogenesis always precedes hairy-root formation, and after 15 days, when roots develop at wounded sites, the stem with the normal root is cleaved below the hairy root zone. Transgenic roots and nodules co-transformed with a binary vector can be easily identified using a reporter gene. This procedure, in addition to inducing robust transgenic hairy roots that are susceptible to being nodulated by rhizobia and to fixing nitrogen efficiently, sets the foundation for a high-throughput functional genomics approach on the study of root biology and root–microbe interactions. This protocol can be completed within 30 days. |
| doi_str_mv | 10.1038/nprot.2007.259 |
| format | Article |
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Phaseolus vulgaris
(common bean) and other
Phaseolus
spp. Hairy roots rapidly develop after wounding young plantlets with
Agrobacterium rhizogenes
, at the cotyledon node, and keeping the plants in high-humidity conditions. Callogenesis always precedes hairy-root formation, and after 15 days, when roots develop at wounded sites, the stem with the normal root is cleaved below the hairy root zone. Transgenic roots and nodules co-transformed with a binary vector can be easily identified using a reporter gene. This procedure, in addition to inducing robust transgenic hairy roots that are susceptible to being nodulated by rhizobia and to fixing nitrogen efficiently, sets the foundation for a high-throughput functional genomics approach on the study of root biology and root–microbe interactions. This protocol can be completed within 30 days.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/nprot.2007.259</identifier><identifier>PMID: 17641650</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>Agrobacterium rhizogenes ; Agrobacterium tumefaciens ; Analysis ; Analytical Chemistry ; Animal Feed ; Animals ; Bacteria ; Biological Techniques ; Biomedical and Life Sciences ; Computational Biology/Bioinformatics ; Cultivars ; Fabaceae - microbiology ; Food ; Gene Expression Regulation, Bacterial ; Genetic engineering ; Genetic transformation ; Genomics ; Humans ; Humidity ; Legumes ; Life Sciences ; Methods ; Microarrays ; Nitrogen ; Organic Chemistry ; Phaseolus ; Phaseolus - genetics ; Phaseolus - microbiology ; Phaseolus vulgaris ; Plant Diseases - microbiology ; Plant Roots - microbiology ; Protocol ; Reproducibility of Results ; Rhizobium - genetics ; Root zone ; Roots ; Transformation, Bacterial ; Ventilation</subject><ispartof>Nature protocols, 2007-07, Vol.2 (7), p.1819-1824</ispartof><rights>Springer Nature Limited 2007</rights><rights>COPYRIGHT 2007 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Jul 2007</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c635t-1eaf1dd6a6d140aa0104e51288b8d389992d7975a7a4382efa60749381bcbcf93</citedby><cites>FETCH-LOGICAL-c635t-1eaf1dd6a6d140aa0104e51288b8d389992d7975a7a4382efa60749381bcbcf93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nprot.2007.259$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nprot.2007.259$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27903,27904,41467,42536,51298</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17641650$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Estrada-Navarrete, Georgina</creatorcontrib><creatorcontrib>Alvarado-Affantranger, Xochitl</creatorcontrib><creatorcontrib>Olivares, Juan-Elías</creatorcontrib><creatorcontrib>Guillén, Gabriel</creatorcontrib><creatorcontrib>Díaz-Camino, Claudia</creatorcontrib><creatorcontrib>Campos, Francisco</creatorcontrib><creatorcontrib>Quinto, Carmen</creatorcontrib><creatorcontrib>Gresshoff, Peter M</creatorcontrib><creatorcontrib>Sanchez, Federico</creatorcontrib><title>Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>This transformation procedure generates, with high efficiency (70–90%), hairy roots in cultivars, landraces and accessions of
Phaseolus vulgaris
(common bean) and other
Phaseolus
spp. Hairy roots rapidly develop after wounding young plantlets with
Agrobacterium rhizogenes
, at the cotyledon node, and keeping the plants in high-humidity conditions. Callogenesis always precedes hairy-root formation, and after 15 days, when roots develop at wounded sites, the stem with the normal root is cleaved below the hairy root zone. Transgenic roots and nodules co-transformed with a binary vector can be easily identified using a reporter gene. This procedure, in addition to inducing robust transgenic hairy roots that are susceptible to being nodulated by rhizobia and to fixing nitrogen efficiently, sets the foundation for a high-throughput functional genomics approach on the study of root biology and root–microbe interactions. This protocol can be completed within 30 days.</description><subject>Agrobacterium rhizogenes</subject><subject>Agrobacterium tumefaciens</subject><subject>Analysis</subject><subject>Analytical Chemistry</subject><subject>Animal Feed</subject><subject>Animals</subject><subject>Bacteria</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Computational Biology/Bioinformatics</subject><subject>Cultivars</subject><subject>Fabaceae - microbiology</subject><subject>Food</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genetic engineering</subject><subject>Genetic transformation</subject><subject>Genomics</subject><subject>Humans</subject><subject>Humidity</subject><subject>Legumes</subject><subject>Life Sciences</subject><subject>Methods</subject><subject>Microarrays</subject><subject>Nitrogen</subject><subject>Organic Chemistry</subject><subject>Phaseolus</subject><subject>Phaseolus - genetics</subject><subject>Phaseolus - microbiology</subject><subject>Phaseolus vulgaris</subject><subject>Plant Diseases - microbiology</subject><subject>Plant Roots - microbiology</subject><subject>Protocol</subject><subject>Reproducibility of Results</subject><subject>Rhizobium - genetics</subject><subject>Root zone</subject><subject>Roots</subject><subject>Transformation, Bacterial</subject><subject>Ventilation</subject><issn>1754-2189</issn><issn>1750-2799</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkl1rFDEYhQdRbK3eeikBQRDdaT5mJsnlslgtFBWteBkymTfblJlkTTJg_fVm24W6siK5SEieczh5OVX1nOCaYCZO_SaGXFOMeU1b-aA6JrzFC8qlfHh7bhaUCHlUPUnpGuOGs44_ro4I7xrStfi4gjOd8lsE1jrjwGek_YAiFNdhNq4fAa3BQ3YG5ah9siFOOrvgUbDo85VOEMY5obTZ1Ki_Qct1DL02GaKbJxSv3K-wlaen1SOrxwTPdvtJ9e3s3eXqw-Li0_vz1fJiYTrW5gUBbckwdLobSIO1xgQ30BIqRC8GJqSUdOCSt5rrhgkKVneYN5IJ0pveWMlOqld3viX_jxlSVpNLBsZRewhzUhzzTjD8f5BiynBDRQFf_gVehzn68glFGOWUSYn5PbXWIyjnbSjTMltLtSSC4qbk3HrVB6iyBpicCR6sK_d7gtd7gsJk-JnXek5JnX_9ss---Te7vPy--ngwiokhpQhWbaKbdLxRBKttsdRtsdS2WKoUqwhe7OYw9xMM9_iuSQU4vQNSefJriH8M6rDlbxX-1qA</recordid><startdate>20070701</startdate><enddate>20070701</enddate><creator>Estrada-Navarrete, Georgina</creator><creator>Alvarado-Affantranger, Xochitl</creator><creator>Olivares, Juan-Elías</creator><creator>Guillén, Gabriel</creator><creator>Díaz-Camino, Claudia</creator><creator>Campos, Francisco</creator><creator>Quinto, Carmen</creator><creator>Gresshoff, Peter M</creator><creator>Sanchez, Federico</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ATWCN</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7QL</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>20070701</creationdate><title>Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes</title><author>Estrada-Navarrete, Georgina ; Alvarado-Affantranger, Xochitl ; Olivares, Juan-Elías ; Guillén, Gabriel ; Díaz-Camino, Claudia ; Campos, Francisco ; Quinto, Carmen ; Gresshoff, Peter M ; Sanchez, Federico</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c635t-1eaf1dd6a6d140aa0104e51288b8d389992d7975a7a4382efa60749381bcbcf93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Agrobacterium rhizogenes</topic><topic>Agrobacterium tumefaciens</topic><topic>Analysis</topic><topic>Analytical Chemistry</topic><topic>Animal Feed</topic><topic>Animals</topic><topic>Bacteria</topic><topic>Biological Techniques</topic><topic>Biomedical and Life Sciences</topic><topic>Computational Biology/Bioinformatics</topic><topic>Cultivars</topic><topic>Fabaceae - 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Academic</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Estrada-Navarrete, Georgina</au><au>Alvarado-Affantranger, Xochitl</au><au>Olivares, Juan-Elías</au><au>Guillén, Gabriel</au><au>Díaz-Camino, Claudia</au><au>Campos, Francisco</au><au>Quinto, Carmen</au><au>Gresshoff, Peter M</au><au>Sanchez, Federico</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2007-07-01</date><risdate>2007</risdate><volume>2</volume><issue>7</issue><spage>1819</spage><epage>1824</epage><pages>1819-1824</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>This transformation procedure generates, with high efficiency (70–90%), hairy roots in cultivars, landraces and accessions of
Phaseolus vulgaris
(common bean) and other
Phaseolus
spp. Hairy roots rapidly develop after wounding young plantlets with
Agrobacterium rhizogenes
, at the cotyledon node, and keeping the plants in high-humidity conditions. Callogenesis always precedes hairy-root formation, and after 15 days, when roots develop at wounded sites, the stem with the normal root is cleaved below the hairy root zone. Transgenic roots and nodules co-transformed with a binary vector can be easily identified using a reporter gene. This procedure, in addition to inducing robust transgenic hairy roots that are susceptible to being nodulated by rhizobia and to fixing nitrogen efficiently, sets the foundation for a high-throughput functional genomics approach on the study of root biology and root–microbe interactions. This protocol can be completed within 30 days.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>17641650</pmid><doi>10.1038/nprot.2007.259</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| issn | 1754-2189 1750-2799 |
| language | eng |
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| source | MEDLINE; Nature; Springer Nature - Complete Springer Journals |
| subjects | Agrobacterium rhizogenes Agrobacterium tumefaciens Analysis Analytical Chemistry Animal Feed Animals Bacteria Biological Techniques Biomedical and Life Sciences Computational Biology/Bioinformatics Cultivars Fabaceae - microbiology Food Gene Expression Regulation, Bacterial Genetic engineering Genetic transformation Genomics Humans Humidity Legumes Life Sciences Methods Microarrays Nitrogen Organic Chemistry Phaseolus Phaseolus - genetics Phaseolus - microbiology Phaseolus vulgaris Plant Diseases - microbiology Plant Roots - microbiology Protocol Reproducibility of Results Rhizobium - genetics Root zone Roots Transformation, Bacterial Ventilation |
| title | Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes |
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